Clinical Significance Of DcR3 Gene Amplification And Expression In Primary Lung Cancer

【Objective】Detect overexpression and amplification of DcR3 gene in primary lung cancer and evaluate its clinical significance.【Methods】1. Establish a robust, specific, and sensitive ELISA with DcR3 monoclonal antibody to detect soluble DcR3 in sera of patients with 26 primary lung cancers, 20 inflammatory diseases, and 29 healthy individuals, and determine the differences of sera DcR3 among primary lung cancers, inflammatory diseases and healthy individuals.2. Study DcR3 expression in 26 primary lung cancer masses with immunohistochemistry to identify the source of elevated serum DcR3.3. Extract genomic DNA form 26 fresh primary lung cancer tissues and peripheral blood leukocytes of 5 healthy individuals and establish standard curves of quantitative PCR for DcR3, the target, andβ-globin, the endogenous control respectively. Detect DcR3 gene copy number using relative standard curve method to investigate relationship between DcR3 gene amplification and overexpression.【Results】1. When the concentration and dilution of coating antibody and biotin labeled detection antibody were 6μg/ml and 1:500 respectively, a most optimized calibration curve was generated and the correlation coefficient of it was 0.998 with the standard deviation of 0.028. Regression equation was as follow: y=1.03+0.66 lgx. The inter-assay and intra-assay CVs were less than 10% and the average recovery rate of the assay was 96%.